Direct-fed microbials

ABSTRACT

The invention relates to direct-fed microbials for use in  E. coli  inhibition in animals. More particularly, the invention relates to isolated  Bacillus  strains 101, 235, 77, 177, and 102 and strains having all of the identifying characteristics of these strains, for a use comprising the above-mentioned use. The invention can also be used for treatment of plants and in food processing.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Continuation Application of U.S. patentapplication Ser. No. 15/445,193, filed Feb. 28, 2017, which claimspriority under 35 U.S.C. § 119(e) to U.S. Provisional Application Ser.No. 62/301,438 filed Feb. 29, 2016 and U.S. Provisional Application Ser.No. 62/364,271 filed Jul. 19, 2016, the disclosures of which are herebyexpressly incorporated by reference in their entirety.

FIELD OF THE DISCLOSURE

The invention relates to direct-fed microbials for use in E. coliinhibition in animals. More particularly, the invention relates toisolated Bacillus strains 101, 235, 77, 177, and 102, and strains havingall of the identifying characteristics of these strains, for a usecomprising the above-mentioned use. The invention can also be used fortreatment of plants and in food processing.

BACKGROUND AND SUMMARY OF THE INVENTION

The present invention relates to direct-fed microbial (DFM) compositionsand methods for E. coli inhibition in an animal. An animal'sgastrointestinal tract is constantly challenged by large numbers ofbacteria, viruses, and protozoa found in feed, bedding, and theenvironment. The gastrointestinal tract has a sophisticated system tocounter these potential pathogens consisting of physical, chemical, andimmunological lines of defense. Beneficial bacteria are an importantpart of this system because they provide animals with bacteria thatassist in establishment (or reestablishment) of a normal bacterialprofile, they strengthen the animal's immune system, and they help tofight disease (e.g., disease caused by E. coli in animals). Due to theimportance of preventing and treating E. coli disease in animals, bothto the agricultural industry, and to the human food supply, direct-fedmicrobial strains are needed that inhibit E. coli in animals, such asagricultural animals.

Applicants have developed direct-fed microbials that result in E. coliinhibition in animals. The direct-fed microbials and compositionscomprising the direct-fed microbials described herein offer a commercialbenefit by providing E. coli inhibition in animals, such as agriculturalanimals. In addition, the direct-fed microbial compositions describedherein result in a reduction or elimination in the use of antibioticswhich reduces the overall cost of animal feed.

Methods and compositions are provided for inhibiting E. coli in animals.In various embodiments, the animal can be selected from the groupconsisting of a poultry species, a porcine species, a bovine species, anovine species, an equine species, and a companion animal. In theembodiment where the animal is a poultry species, the poultry speciescan be a broiler chicken. In the embodiment where the animal is aporcine species, the porcine species can be selected from the groupconsisting of a grow finish pig, a nursery pig, a sow, and a breedingstock pig.

In various embodiments, the compositions for use in the methodsdescribed herein can be a commercial package, a feed additive for ananimal feed composition, an additive for the drinking water of ananimal, or an animal feed composition (e.g., a complete feed), eachcomprising an isolated Bacillus strain selected from the groupconsisting of Bacillus strain 101 (NRRL No. B-67218), Bacillus strain235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274), Bacillusstrain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), astrain having all of the identifying characteristics of Bacillus strain101 (NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof.

In one embodiment of the methods described herein, a method is providedof feeding an animal. The method comprises the step of administering tothe animal a feed composition or drinking water comprising an effectiveamount of an additive comprising an isolated Bacillus strain selectedfrom the group consisting of Bacillus strain 101 (NRRL No. B-67218),Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102(NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof, wherein the Bacillus strain causes E. coliinhibition in the animal.

In another embodiment of the methods described herein, a method isprovided of controlling a detrimental effect of E. coli. The methodcomprises the steps of administering to an animal a feed composition ordrinking water comprising an effective amount of an additive comprisingan isolated Bacillus strain selected from the group consisting ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), Bacillus strain 77 (NRRL No. B-67274), Bacillus strain 177(NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof, and controlling thedetrimental effect of E. coli in the animal.

The following clauses, and combinations thereof, provide variousadditional illustrative aspects of the invention described herein. Thevarious embodiments described in any other section of this patentapplication, including the section titled “DETAILED DESCRIPTION OFILLUSTRATIVE EMBODIMENTS” and the EXAMPLES are applicable to any of thefollowing embodiments of the invention described in the numbered clausesbelow.

1. A method of feeding an animal, the method comprising the step ofadministering to the animal a feed composition or drinking watercomprising an effective amount of an additive comprising an isolatedBacillus strain selected from the group consisting of Bacillus strain101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillusstrain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275),Bacillus strain 102 (NRRL No. B-67276), a strain having all of theidentifying characteristics of Bacillus strain 101 (NRRL No. B-67218), astrain having all of the identifying characteristics of Bacillus strain235 (NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof, wherein the Bacillus strain causes E. coliinhibition in the animal.

2. The method of clause 1 wherein the animal is selected from the groupconsisting of a poultry species, a porcine species, a bovine species, anovine species, an equine species, and a companion animal.

3. The method of clause 2 wherein the poultry species is a broilerchicken.

4. The method of any one of clauses 1 to 3 wherein the E. coliinhibition prevents E. coli disease in the animal.

5. The method of any one of clauses 1 to 3 wherein the E. coliinhibition reduces E. coli disease in the animal.

6. The method of clause 2 wherein the animal is a porcine species andthe porcine species is selected from the group consisting of a growfinish pig, a nursery pig, a sow, and a breeding stock pig.

7. The method of any one of clauses 1 to 6 wherein the Bacillus strainproduces an enzyme selected from the group consisting of anα-galactosidase, a protease, a lipase, an amylase, a xylanase, acellulase, and combinations thereof.

8. The method of any one of clauses 1 to 7 further comprising the stepof administering to the animal another bacterial strain selected fromthe group consisting of another Bacillus strain, a lactic acid bacterialstrain, and combinations thereof.

9. The method of any one of clauses 1 to 8 wherein the strainadministered is Bacillus strain 101 (NRRL No. B-67218) or a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218).

10. The method of any one of clauses 1 to 8 wherein the strainadministered is Bacillus strain 235 (NRRL No. B-67219) or a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219).

11. The method of any one of clauses 1 to 8 wherein Bacillus strain 101(NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in a single composition.

12. The method of any one of clauses 1 to 8 wherein Bacillus strain 101(NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in separate compositions.

13. The method of any one of clauses 1 to 12 wherein the Bacillus strainis administered in the feed composition at a dose of about 1.0×10³CFU/gram of the feed composition to about 5.0×10¹² CFU/gram of the feedcomposition.

14. The method of any one of clauses 1 to 12 wherein the Bacillus strainis administered in the feed composition at a dose of about 1.0×10³CFU/gram of the feed composition to about 1.0×10⁷ CFU/gram of the feedcomposition.

15. The method of any one of clauses 1 to 14 wherein the Bacillus strainis administered in the feed composition at a dose greater than about7.0×10⁴ CFU/gram of the feed composition.

16. The method of any one of clauses 1 to 15 wherein the Bacillus strainis administered in the feed composition at a dose of about 7.3×10⁴CFU/gram of the feed composition.

17. The method of any one of clauses 1 to 16 wherein the Bacillus strainis isolated from a high performing grow finish pig.

18. The method of any one of clauses 1 to 17 further comprising the stepof administering an antibiotic to the animal.

19. The method of any one of clauses 1 to 18 further comprising the stepof administering to the animal an enzyme selected from the groupconsisting of a galactosidase, a protease, a lipase, an amylase, ahemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, aphytase, and combinations thereof.

20. The method of clause 19 wherein the enzyme is an NSPase or aphytase.

21. The method of any one of clauses 1 to 20 wherein the microbialbalance in the animal is maintained.

22. The method of clause 2 wherein the animal is a companion animal.

23. The method of clause 22 wherein the animal is a canine species or afeline species.

24. The method of clause 2 wherein the animal is a sow and the Bacillusstrain is administered during lactation.

25. The method of clause 2 wherein the animal is a sow and the Bacillusstrain is administered during gestation.

26. The method of any one of clauses 1 to 25 wherein the feedcomposition is administered daily to the animal.

27. The method of clause 1 wherein the animal is selected from the groupconsisting of a chicken, a pig, a horse, a pony, a cow, a turkey, agoat, a sheep, a quail, a pheasant, an ostrich, a duck, a fish, acrustacean, and combinations thereof.

28. A method of controlling a detrimental effect of E. coli, the methodcomprising the steps of administering to an animal a feed composition ordrinking water comprising an effective amount of an additive comprisingan isolated Bacillus strain selected from the group consisting ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), Bacillus strain 77 (NRRL No. B-67274), Bacillus strain 177(NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof, and controlling thedetrimental effect of E. coli in the animal.

29. The method of clause 28 wherein the animal is selected from thegroup consisting of a poultry species, a porcine species, a bovinespecies, an ovine species, an equine species, and a companion animal.

30. The method of clause 29 wherein the poultry species is a broilerchicken.

31. The method of any one of clauses 28 to 30 wherein controlling thedetrimental effect of E. coli comprises inhibiting E. coli disease inthe animal.

32. The method of any one of clauses 28 to 30 wherein controlling thedetrimental effect of E. coli comprises reducing E. coli disease in theanimal.

33. The method of clause 29 wherein the animal is a porcine species andthe porcine species is selected from the group consisting of a growfinish pig, a nursery pig, a sow, and a breeding stock pig.

34. The method of any one of clauses 28 to 33 wherein the Bacillusstrain produces an enzyme selected from the group consisting of anα-galactosidase, a protease, a lipase, an amylase, a xylanase, acellulase, and combinations thereof.

35. The method of any one of clauses 28 to 34 further comprising thestep of administering to the animal another bacterial strain selectedfrom the group consisting of another Bacillus strain, a lactic acidbacterial strain, and combinations thereof.

36. The method of any one of clauses 28 to 35 wherein the strainadministered is Bacillus strain 101 (NRRL No. B-67218) or a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218).

37. The method of any one of clauses 28 to 35 wherein the strainadministered is Bacillus strain 235 (NRRL No. B-67219) or a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219).

38. The method of any one of clauses 28 to 35 wherein Bacillus strain101 (NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in a single composition.

39. The method of any one of clauses 28 to 35 wherein Bacillus strain101 (NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in separate compositions.

40. The method of any one of clauses 28 to 39 wherein the Bacillusstrain is administered in the feed composition at a dose of about1.0×10³ CFU/gram of the feed composition to about 5.0×10¹² CFU/gram ofthe feed composition.

41. The method of any one of clauses 28 to 40 wherein the Bacillusstrain is administered in the feed composition at a dose of about1.0×10³ CFU/gram of the feed composition to about 1.0×10⁷ CFU/gram ofthe feed composition.

42. The method of any one of clauses 28 to 41 wherein the Bacillusstrain is administered in the feed composition at a dose greater thanabout 7.0×10⁴ CFU/gram of the feed composition.

43. The method of any one of clauses 28 to 42 wherein the Bacillusstrain is administered in the feed composition at a dose of about7.3×10⁴ CFU/gram of the feed composition.

44. The method of any one of clauses 28 to 43 wherein the Bacillusstrain is isolated from a high performing grow finish pig.

45. The method of any one of clauses 28 to 44 further comprising thestep of administering an antibiotic to the animal.

46. The method of any one of clauses 28 to 45 further comprising thestep of administering to the animal an enzyme selected from the groupconsisting of a galactosidase, a protease, a lipase, an amylase, ahemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, aphytase, and combinations thereof.

47. The method of clause 46 wherein the enzyme is an NSPase or aphytase.

48. The method of any one of clauses 28 to 47 wherein controlling thedetrimental effect of E. coli comprises maintaining the microbialbalance in the animal.

49. The method of clause 29 wherein the animal is a companion animal.

50. The method of clause 49 wherein the animal is a canine species or afeline species.

51. The method of clause 29 wherein the animal is a sow and the Bacillusstrain is administered during lactation.

52. The method of clause 29 wherein the animal is a sow and the Bacillusstrain is administered during gestation.

53. The method of any one of clauses 28 to 52 wherein the feedcomposition is administered daily to the animal.

54. The method of clause 28 wherein the animal is selected from thegroup consisting of a chicken, a pig, a horse, a pony, a cow, a turkey,a goat, a sheep, a quail, a pheasant, an ostrich, a duck, a fish, acrustacean, and combinations thereof.

55. A commercial package comprising an isolated Bacillus strain selectedfrom the group consisting of Bacillus strain 101 (NRRL No. B-67218),Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102(NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

56. A feed additive for an animal feed comprising an isolated Bacillusstrain selected from the group consisting of Bacillus strain 101 (NRRLNo. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77(NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillusstrain 102 (NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

57. An additive for the drinking water of an animal comprising anisolated Bacillus strain selected from the group consisting of Bacillusstrain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219),Bacillus strain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No.B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all ofthe identifying characteristics of Bacillus strain 101 (NRRL No.B-67218), a strain having all of the identifying characteristics ofBacillus strain 235 (NRRL No. B-67219), a strain having all of theidentifying characteristics of Bacillus strain 77 (NRRL No. B-67274), astrain having all of the identifying characteristics of Bacillus strain177 (NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

58. An animal feed composition comprising an isolated Bacillus strainselected from the group consisting of Bacillus strain 101 (NRRL No.B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77(NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillusstrain 102 (NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

59. The commercial package, feed additive, feed composition, or additivefor the drinking water of the animal of any one of clauses 55 to 58wherein the Bacillus strain causes an effect selected from the groupconsisting of preventing E. coli disease, reducing E. coli disease,maintaining the microbial balance of the animal, and combinationsthereof.

60. The commercial package, feed additive, feed composition, or additivefor the drinking water of the animal of any one of clauses 55 to 59,wherein the Bacillus strain reduces E. coli disease in the animal.

61. The feed additive or additive for the drinking water of the animalof clause 56 or 57 in the form of a concentrate.

62. The feed additive or additive for the drinking water of the animalof clause 56 or 57 in the form of a superconcentrate.

63. The feed additive, feed composition, or additive for the drinkingwater of the animal of any one of clauses 56 to 62 in dry form.

64. The feed composition of clause 63 in pelleted form.

65. The commercial package of clause 55 wherein the strains are in aform selected from the group consisting of a powder, a liquid, and apellet form.

66. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 65further comprising a carrier for the Bacillus strains.

67. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 66 wherein thecarrier is selected from the group consisting of a bran, rice hulls, asalt, a dextrin, and combinations thereof.

68. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 67in a bag.

69. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 68 wherein the bag isa plastic bag.

70. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 69further comprising instructions for use of one or more of the Bacillusstrains.

71. The commercial package, feed additive, feed composition, or additivefor the drinking water of the animal of any one of clauses 68 to 70 in a20-pound bag.

72. The commercial package, feed additive, feed composition, or additivefor the drinking water of the animal of any one of clauses 68 to 70 in a50-pound bag.

73. The feed additive or additive for the drinking water of the animalof any one of clauses 56, 57, 59 to 63, or 66 to 72 in powder form.

74. The feed additive or additive for the drinking water of the animalof any one of clauses 56, 57, 59 to 60, or 68 to 70 in liquid form.

75. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 74in a container for commercial use.

76. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 75 wherein thecontainer comprises plastic.

77. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 75 wherein thecontainer comprises paper.

78. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 77further comprising a binder.

79. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 78 wherein the binderis selected from the group consisting of clay, yeast cell wallcomponents, aluminum silicate, and glucan, or combinations thereof.

80. A method of feeding an animal, the method comprising the step ofadministering to the animal a feed composition or drinking watercomprising an effective amount of an additive comprising an isolatedBacillus strain selected from the group consisting of Bacillus strain101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillusstrain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275),Bacillus strain 102 (NRRL No. B-67276), a strain having all of theidentifying characteristics of Bacillus strain 101 (NRRL No. B-67218), astrain having all of the identifying characteristics of Bacillus strain235 (NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

81. The method of clause 80 wherein the animal is selected from thegroup consisting of a poultry species, a porcine species, a bovinespecies, an ovine species, an equine species, and a companion animal.

82. The method of clause 81 wherein the poultry species is a broilerchicken.

83. The method of any one of clauses 80 to 82 wherein the strain causesE. coli inhibition which prevents E. coli disease in the animal.

84. The method of any one of clauses 80 to 82 wherein the strain causesE. coli inhibition which reduces E. coli disease in the animal.

85. The method of clause 81 wherein the animal is a porcine species andthe porcine species is selected from the group consisting of a growfinish pig, a nursery pig, a sow, and a breeding stock pig.

86. The method of any one of clauses 80 to 85 wherein the Bacillusstrain produces an enzyme selected from the group consisting of anα-galactosidase, a protease, a lipase, an amylase, a xylanase, acellulase, and combinations thereof.

87. The method of any one of clauses 80 to 86 further comprising thestep of administering to the animal another bacterial strain selectedfrom the group consisting of another Bacillus strain, a lactic acidbacterial strain, and combinations thereof.

88. The method of any one of clauses 80 to 87 wherein the strainadministered is Bacillus strain 101 (NRRL No. B-67218) or a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218).

89. The method of any one of clauses 80 to 87 wherein the strainadministered is Bacillus strain 235 (NRRL No. B-67219) or a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219).

90. The method of any one of clauses 80 to 87 wherein Bacillus strain101 (NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in a single composition.

91. The method of any one of clauses 80 to 87 wherein Bacillus strain101 (NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in separate compositions.

92. The method of any one of clauses 80 to 91 wherein the Bacillusstrain is administered in the feed composition at a dose of about1.0×10³ CFU/gram of the feed composition to about 5.0×10¹² CFU/gram ofthe feed composition.

93. The method of any one of clauses 80 to 92 wherein the Bacillusstrain is administered in the feed composition at a dose of about1.0×10³ CFU/gram of the feed composition to about 1.0×10⁷ CFU/gram ofthe feed composition.

94. The method of any one of clauses 80 to 93 wherein the Bacillusstrain is administered in the feed composition at a dose greater thanabout 7.0×10⁴ CFU/gram of the feed composition.

95. The method of any one of clauses 80 to 94 wherein the Bacillusstrain is administered in the feed composition at a dose of about7.3×10⁴ CFU/gram of the feed composition.

96. The method of any one of clauses 80 to 95 wherein the Bacillusstrain is isolated from a high performing grow finish pig.

97. The method of any one of clauses 80 to 96 further comprising thestep of administering an antibiotic to the animal.

98. The method of any one of clauses 80 to 97 further comprising thestep of administering to the animal an enzyme selected from the groupconsisting of a galactosidase, a protease, a lipase, an amylase, ahemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, aphytase, and combinations thereof.

99. The method of clause 98 wherein the enzyme is an NSPase or aphytase.

100. The method of any one of clauses 80 to 99 wherein the microbialbalance in the animal is maintained.

101. The method of clause 81 wherein the animal is a companion animal.

102. The method of clause 101 wherein the animal is a canine species ora feline species.

103. The method of clause 81 wherein the animal is a sow and theBacillus strain is administered during lactation.

104. The method of clause 81 wherein the animal is a sow and theBacillus strain is administered during gestation.

105. The method of any one of clauses 80 to 104 wherein the feedcomposition is administered daily to the animal.

106. The method of clause 80 wherein the animal is selected from thegroup consisting of a chicken, a pig, a horse, a pony, a cow, a turkey,a goat, a sheep, a quail, a pheasant, an ostrich, a duck, a fish, acrustacean, and combinations thereof.

107. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein the strain administered is Bacillus strain 77 (NRRL No. B-67274)or a strain having all of the identifying characteristics of Bacillusstrain 77 (NRRL No. B-67274).

108. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein the strain administered is Bacillus strain 177 (NRRL No.B-67275) or a strain having all of the identifying characteristics ofBacillus strain 177 (NRRL No. B-67275).

109. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein the strain administered is Bacillus strain 102 (NRRL No.B-67276), or a strain having all of the identifying characteristics ofBacillus strain 102 (NRRL No. B-67276).

110. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein isolated Bacillus strains comprising Bacillus strains 101 (NRRLNo. B-67218), 235 (NRRL No. B-67219), 77 (NRRL No. B-67274), 177 (NRRLNo. B-67275), and 102 (NRRL No. B-67276) are administered in combinationin separate compositions.

111. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein isolated Bacillus strains comprising Bacillus strains 101 (NRRLNo. B-67218), 235 (NRRL No. B-67219), 77 (NRRL No. B-67274), 177 (NRRLNo. B-67275), and 102 (NRRL No. B-67276) are administered in combinationin a single composition.

112. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein isolated Bacillus strains comprising Bacillus strains 101 (NRRLNo. B-67218), 235 (NRRL No. B-67219), 177 (NRRL No. B-67275), and 102(NRRL No. B-67276) are administered in combination in separatecompositions.

113. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein isolated Bacillus strains comprising Bacillus strains 101 (NRRLNo. B-67218), 235 (NRRL No. B-67219), 177 (NRRL No. B-67275), and 102(NRRL No. B-67276) are administered in combination in a singlecomposition.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

Methods and compositions are provided for inhibiting E. coli in animals,such as agricultural animals. In various embodiments, the compositionsfor use in the methods described herein can be a commercial package, afeed additive for an animal feed composition, an additive for thedrinking water of an animal, or an animal feed composition (e.g., acomplete feed), each comprising an isolated Bacillus strain selectedfrom the group consisting of Bacillus strain 101 (NRRL No. B-67218),Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102(NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

In one embodiment of the methods described herein, a method is providedof feeding an animal. The method comprises the step of administering tothe animal a feed composition or drinking water comprising an effectiveamount of an additive comprising an isolated Bacillus strain selectedfrom the group consisting of Bacillus strain 101 (NRRL No. B-67218),Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102(NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof, wherein the Bacillus strain causes E. coliinhibition in the animal.

In another embodiment of the methods described herein, a method isprovided of controlling a detrimental effect of E. coli. The methodcomprises the steps of administering to an animal a feed composition ordrinking water comprising an effective amount of an additive comprisingan isolated Bacillus strain selected from the group consisting ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), Bacillus strain 77 (NRRL No. B-67274), Bacillus strain 177(NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof, and controlling thedetrimental effect of E. coli in the animal.

The following clauses, and combinations thereof, provide variousadditional illustrative aspects of the invention described herein. Thevarious embodiments described in this section titled “DETAILEDDESCRIPTION OF ILLUSTRATIVE EMBODIMENTS” are applicable to any of thefollowing embodiments of the invention described in the numbered clausesbelow.

1. A method of feeding an animal, the method comprising the step ofadministering to the animal a feed composition or drinking watercomprising an effective amount of an additive comprising an isolatedBacillus strain selected from the group consisting of Bacillus strain101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillusstrain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275),Bacillus strain 102 (NRRL No. B-67276), a strain having all of theidentifying characteristics of Bacillus strain 101 (NRRL No. B-67218), astrain having all of the identifying characteristics of Bacillus strain235 (NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof, wherein the Bacillus strain causes E. coliinhibition in the animal.

2. The method of clause 1 wherein the animal is selected from the groupconsisting of a poultry species, a porcine species, a bovine species, anovine species, an equine species, and a companion animal.

3. The method of clause 2 wherein the poultry species is a broilerchicken.

4. The method of any one of clauses 1 to 3 wherein the E. coliinhibition prevents E. coli disease in the animal.

5. The method of any one of clauses 1 to 3 wherein the E. coliinhibition reduces E. coli disease in the animal.

6. The method of clause 2 wherein the animal is a porcine species andthe porcine species is selected from the group consisting of a growfinish pig, a nursery pig, a sow, and a breeding stock pig.

7. The method of any one of clauses 1 to 6 wherein the Bacillus strainproduces an enzyme selected from the group consisting of anα-galactosidase, a protease, a lipase, an amylase, a xylanase, acellulase, and combinations thereof.

8. The method of any one of clauses 1 to 7 further comprising the stepof administering to the animal another bacterial strain selected fromthe group consisting of another Bacillus strain, a lactic acid bacterialstrain, and combinations thereof.

9. The method of any one of clauses 1 to 8 wherein the strainadministered is Bacillus strain 101 (NRRL No. B-67218) or a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218).

10. The method of any one of clauses 1 to 8 wherein the strainadministered is Bacillus strain 235 (NRRL No. B-67219) or a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219).

11. The method of any one of clauses 1 to 8 wherein Bacillus strain 101(NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in a single composition.

12. The method of any one of clauses 1 to 8 wherein Bacillus strain 101(NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in separate compositions.

13. The method of any one of clauses 1 to 12 wherein the Bacillus strainis administered in the feed composition at a dose of about 1.0×10³CFU/gram of the feed composition to about 5.0×10¹² CFU/gram of the feedcomposition.

14. The method of any one of clauses 1 to 12 wherein the Bacillus strainis administered in the feed composition at a dose of about 1.0×10³CFU/gram of the feed composition to about 1.0×10⁷ CFU/gram of the feedcomposition.

15. The method of any one of clauses 1 to 14 wherein the Bacillus strainis administered in the feed composition at a dose greater than about7.0×10⁴ CFU/gram of the feed composition.

16. The method of any one of clauses 1 to 15 wherein the Bacillus strainis administered in the feed composition at a dose of about 7.3×10⁴CFU/gram of the feed composition.

17. The method of any one of clauses 1 to 16 wherein the Bacillus strainis isolated from a high performing grow finish pig.

18. The method of any one of clauses 1 to 17 further comprising the stepof administering an antibiotic to the animal.

19. The method of any one of clauses 1 to 18 further comprising the stepof administering to the animal an enzyme selected from the groupconsisting of a galactosidase, a protease, a lipase, an amylase, ahemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, aphytase, and combinations thereof.

20. The method of clause 19 wherein the enzyme is an NSPase or aphytase.

21. The method of any one of clauses 1 to 20 wherein the microbialbalance in the animal is maintained.

22. The method of clause 2 wherein the animal is a companion animal.

23. The method of clause 22 wherein the animal is a canine species or afeline species.

24. The method of clause 2 wherein the animal is a sow and the Bacillusstrain is administered during lactation.

25. The method of clause 2 wherein the animal is a sow and the Bacillusstrain is administered during gestation.

26. The method of any one of clauses 1 to 25 wherein the feedcomposition is administered daily to the animal.

27. The method of clause 1 wherein the animal is selected from the groupconsisting of a chicken, a pig, a horse, a pony, a cow, a turkey, agoat, a sheep, a quail, a pheasant, an ostrich, a duck, a fish, acrustacean, and combinations thereof.

28. A method of controlling a detrimental effect of E. coli, the methodcomprising the steps of administering to an animal a feed composition ordrinking water comprising an effective amount of an additive comprisingan isolated Bacillus strain selected from the group consisting ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), Bacillus strain 77 (NRRL No. B-67274), Bacillus strain 177(NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof, and controlling thedetrimental effect of E. coli in the animal.

29. The method of clause 28 wherein the animal is selected from thegroup consisting of a poultry species, a porcine species, a bovinespecies, an ovine species, an equine species, and a companion animal.

30. The method of clause 29 wherein the poultry species is a broilerchicken.

31. The method of any one of clauses 28 to 30 wherein controlling thedetrimental effect of E. coli comprises inhibiting E. coli disease inthe animal.

32. The method of any one of clauses 28 to 30 wherein controlling thedetrimental effect of E. coli comprises reducing E. coli disease in theanimal.

33. The method of clause 29 wherein the animal is a porcine species andthe porcine species is selected from the group consisting of a growfinish pig, a nursery pig, a sow, and a breeding stock pig.

34. The method of any one of clauses 28 to 33 wherein the Bacillusstrain produces an enzyme selected from the group consisting of anα-galactosidase, a protease, a lipase, an amylase, a xylanase, acellulase, and combinations thereof.

35. The method of any one of clauses 28 to 34 further comprising thestep of administering to the animal another bacterial strain selectedfrom the group consisting of another Bacillus strain, a lactic acidbacterial strain, and combinations thereof.

36. The method of any one of clauses 28 to 35 wherein the strainadministered is Bacillus strain 101 (NRRL No. B-67218) or a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218).

37. The method of any one of clauses 28 to 35 wherein the strainadministered is Bacillus strain 235 (NRRL No. B-67219) or a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219).

38. The method of any one of clauses 28 to 35 wherein Bacillus strain101 (NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in a single composition.

39. The method of any one of clauses 28 to 35 wherein Bacillus strain101 (NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in separate compositions.

40. The method of any one of clauses 28 to 39 wherein the Bacillusstrain is administered in the feed composition at a dose of about1.0×10³ CFU/gram of the feed composition to about 5.0×10¹² CFU/gram ofthe feed composition.

41. The method of any one of clauses 28 to 40 wherein the Bacillusstrain is administered in the feed composition at a dose of about1.0×10³ CFU/gram of the feed composition to about 1.0×10⁷ CFU/gram ofthe feed composition.

42. The method of any one of clauses 28 to 41 wherein the Bacillusstrain is administered in the feed composition at a dose greater thanabout 7.0×10⁴ CFU/gram of the feed composition.

43. The method of any one of clauses 28 to 42 wherein the Bacillusstrain is administered in the feed composition at a dose of about7.3×10⁴ CFU/gram of the feed composition.

44. The method of any one of clauses 28 to 43 wherein the Bacillusstrain is isolated from a high performing grow finish pig.

45. The method of any one of clauses 28 to 44 further comprising thestep of administering an antibiotic to the animal.

46. The method of any one of clauses 28 to 45 further comprising thestep of administering to the animal an enzyme selected from the groupconsisting of a galactosidase, a protease, a lipase, an amylase, ahemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, aphytase, and combinations thereof.

47. The method of clause 46 wherein the enzyme is an NSPase or aphytase.

48. The method of any one of clauses 28 to 47 wherein controlling thedetrimental effect of E. coli comprises maintaining the microbialbalance in the animal.

49. The method of clause 29 wherein the animal is a companion animal.

50. The method of clause 49 wherein the animal is a canine species or afeline species.

51. The method of clause 29 wherein the animal is a sow and the Bacillusstrain is administered during lactation.

52. The method of clause 29 wherein the animal is a sow and the Bacillusstrain is administered during gestation.

53. The method of any one of clauses 28 to 52 wherein the feedcomposition is administered daily to the animal.

54. The method of clause 28 wherein the animal is selected from thegroup consisting of a chicken, a pig, a horse, a pony, a cow, a turkey,a goat, a sheep, a quail, a pheasant, an ostrich, a duck, a fish, acrustacean, and combinations thereof.

55. A commercial package comprising an isolated Bacillus strain selectedfrom the group consisting of Bacillus strain 101 (NRRL No. B-67218),Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102(NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

56. A feed additive for an animal feed comprising an isolated Bacillusstrain selected from the group consisting of Bacillus strain 101 (NRRLNo. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77(NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillusstrain 102 (NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

57. An additive for the drinking water of an animal comprising anisolated Bacillus strain selected from the group consisting of Bacillusstrain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219),Bacillus strain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No.B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all ofthe identifying characteristics of Bacillus strain 101 (NRRL No.B-67218), a strain having all of the identifying characteristics ofBacillus strain 235 (NRRL No. B-67219), a strain having all of theidentifying characteristics of Bacillus strain 77 (NRRL No. B-67274), astrain having all of the identifying characteristics of Bacillus strain177 (NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

58. An animal feed composition comprising an isolated Bacillus strainselected from the group consisting of Bacillus strain 101 (NRRL No.B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77(NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillusstrain 102 (NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

59. The commercial package, feed additive, feed composition, or additivefor the drinking water of the animal of any one of clauses 55 to 58wherein the Bacillus strain causes an effect selected from the groupconsisting of preventing E. coli disease, reducing E. coli disease,maintaining the microbial balance of the animal, and combinationsthereof.

60. The commercial package, feed additive, feed composition, or additivefor the drinking water of the animal of any one of clauses 55 to 59,wherein the Bacillus strain reduces E. coli disease in the animal.

61. The feed additive or additive for the drinking water of the animalof clause 56 or 57 in the form of a concentrate.

62. The feed additive or additive for the drinking water of the animalof clause 56 or 57 in the form of a superconcentrate.

63. The feed additive, feed composition, or additive for the drinkingwater of the animal of any one of clauses 56 to 62 in dry form.

64. The feed composition of clause 63 in pelleted form.

65. The commercial package of clause 55 wherein the strains are in aform selected from the group consisting of a powder, a liquid, and apellet form.

66. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 65further comprising a carrier for the Bacillus strains.

67. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 66 wherein thecarrier is selected from the group consisting of a bran, rice hulls, asalt, a dextrin, and combinations thereof.

68. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 67in a bag.

69. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 68 wherein the bag isa plastic bag.

70. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 69further comprising instructions for use of one or more of the Bacillusstrains.

71. The commercial package, feed additive, feed composition, or additivefor the drinking water of the animal of any one of clauses 68 to 70 in a20-pound bag.

72. The commercial package, feed additive, feed composition, or additivefor the drinking water of the animal of any one of clauses 68 to 70 in a50-pound bag.

73. The feed additive or additive for the drinking water of the animalof any one of clauses 56, 57, 59 to 63, or 66 to 72 in powder form.

74. The feed additive or additive for the drinking water of the animalof any one of clauses 56, 57, 59 to 60, or 68 to 70 in liquid form.

75. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 74in a container for commercial use.

76. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 75 wherein thecontainer comprises plastic.

77. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 75 wherein thecontainer comprises paper.

78. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of any one of clauses 55 to 77further comprising a binder.

79. The commercial package, feed additive, additive for the drinkingwater of the animal, or feed composition of clause 78 wherein the binderis selected from the group consisting of clay, yeast cell wallcomponents, aluminum silicate, and glucan, or combinations thereof.

80. A method of feeding an animal, the method comprising the step ofadministering to the animal a feed composition or drinking watercomprising an effective amount of an additive comprising an isolatedBacillus strain selected from the group consisting of Bacillus strain101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillusstrain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275),Bacillus strain 102 (NRRL No. B-67276), a strain having all of theidentifying characteristics of Bacillus strain 101 (NRRL No. B-67218), astrain having all of the identifying characteristics of Bacillus strain235 (NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

81. The method of clause 80 wherein the animal is selected from thegroup consisting of a poultry species, a porcine species, a bovinespecies, an ovine species, an equine species, and a companion animal.

82. The method of clause 81 wherein the poultry species is a broilerchicken.

83. The method of any one of clauses 80 to 82 wherein the strain causesE. coli inhibition which prevents E. coli disease in the animal.

84. The method of any one of clauses 80 to 82 wherein the strain causesE. coli inhibition which reduces E. coli disease in the animal.

85. The method of clause 81 wherein the animal is a porcine species andthe porcine species is selected from the group consisting of a growfinish pig, a nursery pig, a sow, and a breeding stock pig.

86. The method of any one of clauses 80 to 85 wherein the Bacillusstrain produces an enzyme selected from the group consisting of anα-galactosidase, a protease, a lipase, an amylase, a xylanase, acellulase, and combinations thereof.

87. The method of any one of clauses 80 to 86 further comprising thestep of administering to the animal another bacterial strain selectedfrom the group consisting of another Bacillus strain, a lactic acidbacterial strain, and combinations thereof.

88. The method of any one of clauses 80 to 87 wherein the strainadministered is Bacillus strain 101 (NRRL No. B-67218) or a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218).

89. The method of any one of clauses 80 to 87 wherein the strainadministered is Bacillus strain 235 (NRRL No. B-67219) or a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219).

90. The method of any one of clauses 80 to 87 wherein Bacillus strain101 (NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in a single composition.

91. The method of any one of clauses 80 to 87 wherein Bacillus strain101 (NRRL No. B-67218), or a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), and Bacillusstrain 235 (NRRL No. B-67219), or a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), areadministered in combination in separate compositions.

92. The method of any one of clauses 80 to 91 wherein the Bacillusstrain is administered in the feed composition at a dose of about1.0×10³ CFU/gram of the feed composition to about 5.0×10¹² CFU/gram ofthe feed composition.

93. The method of any one of clauses 80 to 92 wherein the Bacillusstrain is administered in the feed composition at a dose of about1.0×10³ CFU/gram of the feed composition to about 1.0×10⁷ CFU/gram ofthe feed composition.

94. The method of any one of clauses 80 to 93 wherein the Bacillusstrain is administered in the feed composition at a dose greater thanabout 7.0×10⁴ CFU/gram of the feed composition.

95. The method of any one of clauses 80 to 94 wherein the Bacillusstrain is administered in the feed composition at a dose of about7.3×10⁴ CFU/gram of the feed composition.

96. The method of any one of clauses 80 to 95 wherein the Bacillusstrain is isolated from a high performing grow finish pig.

97. The method of any one of clauses 80 to 96 further comprising thestep of administering an antibiotic to the animal.

98. The method of any one of clauses 80 to 97 further comprising thestep of administering to the animal an enzyme selected from the groupconsisting of a galactosidase, a protease, a lipase, an amylase, ahemicellulase, an arabinoxylanase, a xylanase, a cellulase, an NSPase, aphytase, and combinations thereof.

99. The method of clause 98 wherein the enzyme is an NSPase or aphytase.

100. The method of any one of clauses 80 to 99 wherein the microbialbalance in the animal is maintained.

101. The method of clause 81 wherein the animal is a companion animal.

102. The method of clause 101 wherein the animal is a canine species ora feline species.

103. The method of clause 81 wherein the animal is a sow and theBacillus strain is administered during lactation.

104. The method of clause 81 wherein the animal is a sow and theBacillus strain is administered during gestation.

105. The method of any one of clauses 80 to 104 wherein the feedcomposition is administered daily to the animal.

106. The method of clause 80 wherein the animal is selected from thegroup consisting of a chicken, a pig, a horse, a pony, a cow, a turkey,a goat, a sheep, a quail, a pheasant, an ostrich, a duck, a fish, acrustacean, and combinations thereof.

107. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein the strain administered is Bacillus strain 77 (NRRL No. B-67274)or a strain having all of the identifying characteristics of Bacillusstrain 77 (NRRL No. B-67274).

108. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein the strain administered is Bacillus strain 177 (NRRL No.B-67275) or a strain having all of the identifying characteristics ofBacillus strain 177 (NRRL No. B-67275).

109. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein the strain administered is Bacillus strain 102 (NRRL No.B-67276), or a strain having all of the identifying characteristics ofBacillus strain 102 (NRRL No. B-67276).

110. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein isolated Bacillus strains comprising Bacillus strains 101 (NRRLNo. B-67218), 235 (NRRL No. B-67219), 77 (NRRL No. B-67274), 177 (NRRLNo. B-67275), and 102 (NRRL No. B-67276) are administered in combinationin separate compositions.

111. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein isolated Bacillus strains comprising Bacillus strains 101 (NRRLNo. B-67218), 235 (NRRL No. B-67219), 77 (NRRL No. B-67274), 177 (NRRLNo. B-67275), and 102 (NRRL No. B-67276) are administered in combinationin a single composition.

112. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein isolated Bacillus strains comprising Bacillus strains 101 (NRRLNo. B-67218), 235 (NRRL No. B-67219), 177 (NRRL No. B-67275), and 102(NRRL No. B-67276) are administered in combination in separatecompositions.

113. The method of any one of clauses 1 to 8, 28 to 35, or 80 to 87wherein isolated Bacillus strains comprising Bacillus strains 101 (NRRLNo. B-67218), 235 (NRRL No. B-67219), 177 (NRRL No. B-67275), and 102(NRRL No. B-67276) are administered in combination in a singlecomposition.

In various embodiments, the animal to which a feed additive, a feedcomposition, or drinking water as described herein is administered canbe selected from the group consisting of a poultry species, a porcinespecies, a bovine species, an ovine species, an equine species, and acompanion animal. In the embodiment where the animal is a companionanimal, the companion animal can be, for example, a canine species or afeline species. In the embodiment where the animal is a porcine species,the porcine species can be selected from the group consisting of a growfinish pig, a nursery pig, a sow, and a breeding stock pig. In variousexemplary embodiments, the animal can be selected from the groupconsisting of a chicken (e.g., a broiler or a layer), a pig, a horse, apony, a cow, a turkey, a goat, a sheep, a quail, a pheasant, an ostrich,a duck, a fish (e.g., a tilapia, a catfish, a flounder, or a salmon), acrustacean (e.g., a shrimp or a crab), and combinations thereof. Inanother embodiment, Bacillus strain 101 (NRRL No. B-67218), Bacillusstrain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274),Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No.B-67276), a strain having all of the identifying characteristics ofBacillus strain 101 (NRRL No. B-67218), a strain having all of theidentifying characteristics of Bacillus strain 235 (NRRL No. B-67219), astrain having all of the identifying characteristics of Bacillus strain77 (NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof, can be used to treat aplant for plant health and can be used to treat the soil, the plantdirectly, or to irrigate the plant.

In any embodiments described herein, the strains can be administered inalone or in any combination, or can be in the form of any compositiondescribed herein so that the strains are alone or in any combination inthe composition described herein. Exemplary combinations include a groupof isolated Bacillus strains comprising Bacillus strains 101 (NRRL No.B-67218), 102 (NRRL No. B-67276), 235 (NRRL No. B-67219), 77 (NRRL No.B-67274), and 177 (NRRL No. B-67275), and a group of isolated Bacillusstrains comprising Bacillus strains 101 (NRRL No. B-67218), 235 (NRRLNo. B-67219), 177 (NRRL No. B-67275), and 102 (NRRL No. B-67276).

In one embodiment, a method of feeding an animal is provided. The methodcomprises the step of administering to the animal a feed composition ordrinking water comprising an effective amount of an additive comprisingan isolated Bacillus strain selected from the group consisting Bacillusstrain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219),Bacillus strain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No.B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all ofthe identifying characteristics of Bacillus strain 101 (NRRL No.B-67218), a strain having all of the identifying characteristics ofBacillus strain 235 (NRRL No. B-67219), a strain having all of theidentifying characteristics of Bacillus strain 77 (NRRL No. B-67274), astrain having all of the identifying characteristics of Bacillus strain177 (NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

In another embodiment, a method of feeding an animal is provided. Themethod comprises the step of administering to the animal a feedcomposition or drinking water comprising an effective amount of anadditive comprising an isolated Bacillus strain selected from the groupconsisting Bacillus strain 101 (NRRL No. B-67218), Bacillus strain 235(NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274), Bacillusstrain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), astrain having all of the identifying characteristics of Bacillus strain101 (NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof, wherein the Bacillusstrain causes E. coli inhibition in the animal.

In one embodiment of the invention, an effective amount of the Bacillusstrain can be administered to inhibit E. coli in the animal. As usedherein, “inhibit E. coli” can mean reducing E. coli disease, preventingE. coli disease, maintaining the normal microbial balance in the animal,reducing the number of detrimental E. coli in the animal, reducing theactivity of E. coli in the animal, or reducing the symptoms of E. colidisease in the animal, or combinations thereof. By “effective amount” ismeant an amount of the Bacillus strain (e.g., strain 101, or 235, or 77,or 177, or 102) capable of E. coli inhibition or capable of controllinga detrimental effect of E. coli, as described below, by any mechanism.

In embodiments described herein wherein the compositions of the presentinvention comprising Bacillus strains 101, and/or 235, and/or 77, and/or177, and/or 102 are administered to an animal, the compositions arepreferably administered to animals orally in a feed composition or indrinking water, but any other effective method of administration knownto those skilled in the art may be utilized. In one illustrativeembodiment, the Bacillus strains 101, and/or 235, and/or 77, and/or 177,and/or 102 are provided in the form of an additive for addition to thedrinking water of an animal.

In another illustrative embodiment, the Bacillus strains 101, and/or235, and/or 77, and/or 177, and/or 102 are provided in the form of afeed additive for addition to a feed composition. The feed compositionmay contain Bacillus strain 101, and/or 235, and/or 77, and/or 177,and/or 102 in a mixture with an animal feed blend, including anyart-recognized animal feed blend or any animal feed blend describedherein. As used herein, “feed composition” or “animal feed composition”means a feed composition comprising Bacillus strain 101, and/or Bacillusstrain 235, and/or Bacillus strain 77, and/or Bacillus strain 177,and/or Bacillus strain 102 in a mixture with an animal feed blend, and,optionally any other components that could be used in a feedcomposition, including other bacterial strains, such as other Bacillusstrains or Lactobacillus strains.

Any animal feed blend, including those known in the art and thosedescribed herein, may be used in accordance with the methods andcompositions described in this patent application, such as rapeseedmeal, cottonseed meal, soybean meal, cornmeal, barley, wheat, silage,and haylage. In various embodiments, the animal feed blend can besupplemented with Bacillus strain 101, and/or Bacillus strain 235,and/or Bacillus strain 77, and/or Bacillus strain 177, and/or Bacillusstrain 102, but other ingredients may optionally be added to the animalfeed blend, including other bacterial strains, such as other Bacillusstrains or Lactobacillus strains.

In various illustrative embodiments, optional ingredients of the animalfeed blend include sugars and complex carbohydrates such as bothwater-soluble and water-insoluble monosaccharides, disaccharides, andpolysaccharides. Other optional ingredients include dried distillersgrain solubles, fat (e.g., crude fat), phosphorous, sodium bicarbonate,limestone, salt, phytate, calcium, sodium, sulfur, magnesium, potassium,copper, iron, manganese, zinc, ash, fish oil, an oil derived from fishmeal, raw seed (e.g., flaxseed), an antioxidant, and starch. In anotherembodiment, minerals may be added in the form of a mineral premix.

Optional amino acid ingredients that may be added to the animal feedblend are arginine, histidine, isoleucine, leucine, lysine, cysteine,methionine, phenylalanine, threonine, tryptophan, valine, tyrosine ethylHCl, alanine, aspartic acid, sodium glutamate, glycine, proline, serine,cysteine ethyl HCl, and analogs, and salts thereof. Vitamins that may beoptionally added are thiamine HCl, riboflavin, pyridoxine HCl, niacin,niacinamide, inositol, choline chloride, calcium pantothenate, biotin,folic acid, ascorbic acid, and vitamins A, B, K, D, E, and the like. Inanother embodiment, vitamins may be added in the form of a vitaminpremix. In yet another embodiment, protein ingredients may be added tothe animal feed blend and include protein obtained from meat meal, bonemeal, or fish meal, liquid or powdered egg, fish solubles, crudeprotein, and the like.

In another illustrative aspect, any medicament ingredients known in theart may be added to the animal feed blend or to an additive for thedrinking water of the animal, such as antibiotics. In variousembodiments, the antibiotic is selected from the group consisting ofampicillin, chloramphenicol, ciprofloxacin, clindamycin, tetracycline,chlortetracycline, Denagard™, BMD™, Carbadox™, Stafac™, erythromycin,levofloxacin, trimethoprim/sulfamethoxazole, trimethoprim, daptomycin,rifampicin, Tylan™, Pulmotil™, vancomycin, and combinations thereof. Inanother embodiment, the animal feed blend, the feed composition, thefeed additive, or the additive for the drinking water of the animal maycontain no antibiotics.

In another illustrative embodiment, one or more enzymes may be added tothe animal feed blend. In various embodiments, the enzymes that may beadded include a galactosidase, a phytase, a protease, a lipase, anamylase, a hemicellulase, an arabinoxylanase, a xylanase, a cellulase,an NSPase, combinations thereof, and any other enzyme that improves theeffectiveness of the feed composition for E. coli inhibition orcontrolling a detrimental effect of E. coli. In yet another embodiment,yeast, fungi (e.g., Aspergillus or Trichoderma), or micronutrients maybe added to the animal feed. Any of the ingredients described above thatare suitable for addition to an additive for the drinking water of theanimal may be added as a component of the additive for the drinkingwater of the animal as described herein.

In various illustrative embodiments, the Bacillus strain (e.g., Bacillusstrain 101, and/or 235, and/or 77, and/or 177, and/or 102), or any otherbacterial strains added in addition to Bacillus strain 101, and/or 235,and/or 77, and/or 177, and/or 102, can be administered in the feedcomposition at a dose of about 1.0×10³ CFU/gram of the feed compositionto about 5.0×10¹² CFU/gram of the feed composition or at a dose of about1.0×10³ CFU/gram of the feed composition to about 1.0×10⁷ CFU/gram ofthe feed composition. In other embodiments, the Bacillus strain (e.g.,Bacillus strain 101, and/or 235, and/or 77, and/or 177, and/or 102) isadministered in the feed composition at a dose greater than about1.0×10³ CFU/gram of the feed composition, at a dose greater than about1.1×10³ CFU/gram of the feed composition, at a dose greater than about1.25×10³ CFU/gram of the feed composition, at a dose greater than about1.5×10³ CFU/gram of the feed composition, at a dose greater than about1.75×10³ CFU/gram of the feed composition, at a dose greater than about1.0×10⁴ CFU/gram of the feed composition, at a dose greater than about2.0×10⁴ CFU/gram of the feed composition, at a dose greater than about3.0×10⁴ CFU/gram of the feed composition, at a dose greater than about4.0×10⁴ CFU/gram of the feed composition, at a dose greater than about5.0×10⁴ CFU/gram of the feed composition, at a dose greater than about6.0×10⁴ CFU/gram of the feed composition, at a dose greater than about7.0×10⁴ CFU/gram of the feed composition, at a dose greater than about8.0×10⁴ CFU/gram of the feed composition, at a dose greater than about1.0×10⁵ CFU/gram of the feed composition, at a dose greater than about1.0×10⁶ CFU/gram of the feed composition, at a dose greater than about1.0×10⁷ CFU/gram of the feed composition, at a dose greater than about1.0×10⁸ CFU/gram of the feed composition, at a dose greater than about1.0×10⁹ CFU/gram of the feed composition, at a dose greater than about1.0×10¹⁰ CFU/gram of the feed composition, at a dose greater than about1.0×10¹¹ CFU/gram of the feed composition, or at a dose greater thanabout 1.0×10¹² CFU/gram of the feed composition. In yet anotherembodiment, the Bacillus strain (e.g., Bacillus strain 101, and/or 235,and/or 77, and/or 177, and/or 102) is administered in the feedcomposition at a dose of about 7.3×10⁴ CFU/gram of the feed composition.In another embodiment, the Bacillus strain (e.g., Bacillus strain 101,and/or 235, and/or 77, and/or 177, and/or 102), or any other bacterialstrains added in addition to Bacillus strain 101, and/or 235, and/or 77,and/or 177, and/or 102, can be administered in the feed composition at adose of about 1.0×10² CFU/gram of the feed composition to about 5.0×10¹²CFU/gram of the feed composition or at a dose of about 1.0×10² CFU/gramof the feed composition to about 1.0×10⁷ CFU/gram of the feedcomposition, or at a dose greater than about 1.0×10² CFU/gram of thefeed composition.

In various embodiments, the Bacillus strain (e.g., Bacillus strain 101,and/or 235, and/or 77, and/or 177, and/or 102) for use in accordancewith the methods and compositions described herein can be selected fromthe group consisting of Bacillus strain 101 (NRRL No. B-67218), Bacillusstrain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274),Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No.B-67276), a strain having all of the identifying characteristics ofBacillus strain 101 (NRRL No. B-67218), a strain having all of theidentifying characteristics of Bacillus strain 235 (NRRL No. B-67219), astrain having all of the identifying characteristics of Bacillus strain77 (NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), and a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276). Bacillus strain MDG 101 and Bacillus strain MDG 235were deposited on Jan. 4, 2016 at the Agricultural Research ServiceCulture Collection (NRRL), National Center for Agricultural UtilizationResearch, Agricultural Research Service, USDA, 1815 North UniversityStreet, Peoria, Ill. 61604-3999, and were given accession numbersB-67218 and B-67219, respectively. Bacillus strain MGL77, Bacillusstrain MGL177, and Bacillus strain MGL102 were deposited on Jun. 7, 2016at the Agricultural Research Service Culture Collection (NRRL), NationalCenter for Agricultural Utilization Research, Agricultural ResearchService, USDA, 1815 North University Street, Peoria, Ill. 61604-3999,and were given accession numbers B-67274, B-67275, and B-67276,respectively. The deposits were made under the provisions of theBudapest Treaty on the International Recognition of the Deposit ofMicroorganisms for the Purposes of Patent Procedure. The NRRL straindesignations are MDG 101, MDG 235, MGL77, MGL177, and MGL102 which areequivalent to Bacillus strain 101, 235, 77, 177, and 102 respectively,as referred to in the application.

Any of these strains can be administered alone or in combination in theform of a feed composition (e.g., a complete feed comprising an animalfeed blend) or drinking water for an animal. In one embodiment, multiplestrains are administered in combination in a single composition. Inanother embodiment, multiple strains are administered in combination inseparate compositions. In one illustrative embodiment, any of thesestrains is isolated from a high performing grow finish pig.

In another embodiment, one or more of the Bacillus strains described inthe preceding paragraphs (e.g., Bacillus strain 101, and/or Bacillusstrain 235, and/or Bacillus strain 77, and/or Bacillus strain 177,and/or Bacillus strain 102) can be administered to the animal along withanother bacterial strain selected from the group consisting of anotherBacillus strain, a lactic acid bacterial strain, and combinationsthereof. In yet another embodiment, one or more of the Bacillus strainsdescribed in the preceding paragraphs (e.g., Bacillus strain 101, and/orBacillus strain 235, and/or Bacillus strain 77, and/or Bacillus strain177, and/or Bacillus strain 102) can be administered to the animal alongwith any other bacterial strain effective to inhibit E. coli in theanimal.

As used herein “a strain having all of the identifying characteristicsof” Bacillus strain 101, or Bacillus strain 235, or Bacillus strain 77,or Bacillus strain 177, or Bacillus strain 102 can be a mutant strainhaving all of the identifying characteristics of Bacillus strain 101, orBacillus strain 235, or Bacillus strain 77, or Bacillus strain 177, orBacillus strain 102 (e.g., a DNA fingerprint based on DNA analysis thatcorresponds to the DNA fingerprint of Bacillus strain 101, or Bacillusstrain 235, or Bacillus strain 77, or Bacillus strain 177, or Bacillusstrain 102, enzyme activities that correspond to Bacillus strain 101, orBacillus strain 235, or Bacillus strain 77, or Bacillus strain 177, orBacillus strain 102, antimicrobial activity that corresponds to Bacillusstrain 101, or Bacillus strain 235, or Bacillus strain or 77, Bacillusstrain or 177, or Bacillus strain 102, antibiotic sensitivity andtolerance profiles that correspond to Bacillus strain 101, or Bacillusstrain 235, or Bacillus strain 77, or Bacillus strain 177, or Bacillusstrain 102, or combinations thereof). In alternate embodiments, themutation can be a natural mutation, or a genetically engineeredmutation. In another embodiment, “a strain having all of the identifyingcharacteristics of” Bacillus strain 101, or Bacillus strain 235, orBacillus strain 77, or Bacillus strain 177, or Bacillus strain 102 canbe a strain, for example, produced by isolating one or more plasmidsfrom Bacillus strain 101, or Bacillus strain 235, or Bacillus strain 77,or Bacillus strain 177, or Bacillus strain 102 and introducing the oneor more plasmids into another bacterium, such as another Bacillusstrain, as long as the one or more plasmids contain DNA that providesthe identifying characteristics of Bacillus strain 101, or Bacillusstrain 235, or Bacillus strain 77, or Bacillus strain 177, or Bacillusstrain 102 (e.g., a DNA fingerprint based on DNA analysis thatcorresponds to the DNA fingerprint of Bacillus strain 101, or Bacillusstrain 235, or Bacillus strain 77, or Bacillus strain 177, or Bacillusstrain 102).

The feed composition or drinking water comprising Bacillus strain 101,and/or 235, and/or 77, and/or 177, and/or 102 may be administered to theanimal for any time period that is effective to inhibit E. coli orcontrol a detrimental effect of E. coli, or combinations thereof. Forexample, in one embodiment the feed composition or drinking water may beprovided to the animal daily. In an alternate embodiment, the feedcomposition or drinking water may be administered to the animal duringlactation and/or during gestation. The time periods for administrationof the feed composition or drinking water described above arenon-limiting examples and it should be appreciated that any time periodor administration schedule determined to be effective to inhibit E. colior control a detrimental effect of E. coli, or combinations thereof, maybe used.

As described herein, one of the method embodiments is a method offeeding an animal by administering to the animal a feed composition ordrinking water comprising an effective amount of an additive comprisingan isolated Bacillus strain selected from the group consisting ofBacillus strain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No.B-67219), Bacillus strain 77 (NRRL No. B-67274), Bacillus strain 177(NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), a strainhaving all of the identifying characteristics of Bacillus strain 101(NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof, wherein the Bacillusstrain controls a detrimental effect of E. coli. Any of the applicableabove-described embodiments can apply to this embodiment.

In this embodiment, “controlling a detrimental effect of E. coli” canmean reducing E. coli disease, preventing E. coli disease, maintainingthe normal microbial balance in the animal, reducing the number ofdetrimental E. coli in the animal, reducing the activity of E. coli inthe animal, or reducing the symptoms of E. coli disease in the animal,or combinations thereof. By “effective amount” is meant an amount of theBacillus strain (e.g., strain 101, or 235, or 77, or 177, or 102)capable of controlling a detrimental effect of E. coli, as describedabove, by any mechanism.

In additional embodiments of the invention, compositions comprisingBacillus strain 101, and/or Bacillus strain 235, and/or Bacillus strain77, and/or Bacillus strain 177, and/or Bacillus strain 102 are provided.In one embodiment, a commercial package is provided comprising anisolated Bacillus strain selected from the group consisting of Bacillusstrain 101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219),Bacillus strain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No.B-67275), Bacillus strain 102 (NRRL No. B-67276), a strain having all ofthe identifying characteristics of Bacillus strain 101 (NRRL No.B-67218), a strain having all of the identifying characteristics ofBacillus strain 235 (NRRL No. B-67219), a strain having all of theidentifying characteristics of Bacillus strain 77 (NRRL No. B-67274), astrain having all of the identifying characteristics of Bacillus strain177 (NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

In another embodiment, a feed additive for an animal feed is providedcomprising an isolated Bacillus strain selected from the groupconsisting of Bacillus strain 101 (NRRL No. B-67218), Bacillus strain235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274), Bacillusstrain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No. B-67276), astrain having all of the identifying characteristics of Bacillus strain101 (NRRL No. B-67218), a strain having all of the identifyingcharacteristics of Bacillus strain 235 (NRRL No. B-67219), a strainhaving all of the identifying characteristics of Bacillus strain 77(NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof.

In yet another embodiment, an additive for the drinking water of ananimal is provided comprising an isolated Bacillus strain selected fromthe group consisting of Bacillus strain 101 (NRRL No. B-67218), Bacillusstrain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274),Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No.B-67276), a strain having all of the identifying characteristics ofBacillus strain 101 (NRRL No. B-67218), a strain having all of theidentifying characteristics of Bacillus strain 235 (NRRL No. B-67219), astrain having all of the identifying characteristics of Bacillus strain77 (NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof.

In yet another illustrative aspect of the invention, an animal feedcomposition is provided comprising an isolated Bacillus strain selectedfrom the group consisting of Bacillus strain 101 (NRRL No. B-67218),Bacillus strain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No.B-67274), Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102(NRRL No. B-67276), a strain having all of the identifyingcharacteristics of Bacillus strain 101 (NRRL No. B-67218), a strainhaving all of the identifying characteristics of Bacillus strain 235(NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof.

In another embodiment an isolated Bacillus strain selected from thegroup consisting of Bacillus strain 101 (NRRL No. B-67218), Bacillusstrain 235 (NRRL No. B-67219), Bacillus strain 77 (NRRL No. B-67274),Bacillus strain 177 (NRRL No. B-67275), Bacillus strain 102 (NRRL No.B-67276), a strain having all of the identifying characteristics ofBacillus strain 101 (NRRL No. B-67218), a strain having all of theidentifying characteristics of Bacillus strain 235 (NRRL No. B-67219), astrain having all of the identifying characteristics of Bacillus strain77 (NRRL No. B-67274), a strain having all of the identifyingcharacteristics of Bacillus strain 177 (NRRL No. B-67275), a strainhaving all of the identifying characteristics of Bacillus strain 102(NRRL No. B-67276), and combinations thereof can be used in maintainingplant health, aquaculture, or for use in food processing.

In one embodiment, the feed additive for addition to an animal feedblend to produce a complete feed composition can be mixed with theanimal feed blend, for example, with an automated micro-nutrientdelivery system, or, for example, by hand-weighing and addition toachieve any of the doses of Bacillus strain 101, Bacillus strain 235,Bacillus strain 77, Bacillus strain 177, and Bacillus strain 102described herein, for administration to the animal in the form of acomplete feed composition. The mixing can also be done by any othersuitable method known in the art for combining direct-fed microbialswith an animal feed blend to obtain a uniform mixture. In variousembodiments, the mixing can be done for any suitable time period (e.g.,about 1 to about 4 minutes). In the embodiment where Bacillus strain101, and/or Bacillus strain 235, and/or Bacillus strain 77, and/orBacillus strain 177, and/or Bacillus strain 102 is in the form of anadditive for the drinking water of the animal, the Bacillus strain 101,and/or Bacillus strain 235, and/or Bacillus strain 77, and/or Bacillusstrain 177, and/or Bacillus strain 102 can be in the form of, forexample, a powder, a liquid, or pellets, and can be mixed with thedrinking water using any suitable method known in the art to achieve anyof the doses of Bacillus strain 101, Bacillus strain 235, Bacillusstrain 77, Bacillus strain 177, or Bacillus strain 102 described herein,for administration to the animal in the drinking water of the animal.Bacillus strain 101, and/or Bacillus strain 235, and/or Bacillus strain77, and/or Bacillus strain 177, and/or Bacillus strain 102 can also befed directly to the animal orally (i.e., by oral insertion) in the formof a powder, a liquid, or a pellet.

In any of the composition embodiments described herein, the Bacillusstrain 101, and/or Bacillus strain 235, and/or Bacillus strain 77,and/or Bacillus strain 177, and/or Bacillus strain 102 can cause aneffect selected from the group consisting of inhibiting E. coli orcontrolling a detrimental effect of E. coli in the animal. These effectsare non-limiting examples of the types of effects Bacillus strain 101,and/or Bacillus strain 235, and/or Bacillus strain 77, and/or Bacillusstrain 177, and/or Bacillus strain 102 can cause.

In one illustrative aspect, the feed additive, additive for the drinkingwater of the animal, or the feed composition can be in the form of acommercial package. In another illustrative embodiment, the feedadditive or additive for the drinking water of the animal, or theBacillus strain 101, and/or Bacillus strain 235, and/or Bacillus strain77, and/or Bacillus strain 177, and/or Bacillus strain 102 in thecommercial package can be in the form of a concentrate (e.g., about1×10⁸ to about 5×10⁹ CFU/g) or a superconcentrate (e.g., about 1×10¹⁰ toabout 5×10¹² CFU/g). In another embodiment, the feed additive, feedcomposition, or additive for the drinking water of the animal, or theBacillus strain 101, and/or Bacillus strain 235, and/or Bacillus strain77, and/or Bacillus strain 177, and/or Bacillus strain 102 in acomposition in a commercial package, can be in a dry form (e.g., apowder), a pelleted form, a liquid form, in the form of a top-dressing,or in the form of a gel, or any other suitable form.

In yet another embodiment, the strains in the form of a commercialpackage can be, for example, in a dry form (e.g., a powder orfreeze-dried form), in a pelleted form, or in a liquid form.

In another illustrative embodiment, the commercial package, feedadditive, additive for the drinking water of the animal, or feedcomposition can further comprise a carrier for the Bacillus strain 101,and/or Bacillus strain 235, and/or Bacillus strain 77, and/or Bacillusstrain 177, and/or Bacillus strain 102. The carrier can be selected fromthe group consisting of a bran, rice hulls, a salt, mineral oil, adextrin (e.g., maltodextrin), whey, sugar, limestone, dried starch,sodium silico aluminate, vegetable oil, and combinations thereof. Inanother embodiment, the carrier can be any suitable carrier known in theart for a direct-fed microbial. In another embodiment, the commercialpackage, feed additive, additive for the drinking water of the animal,or feed composition can further comprise a binder such as clay, yeastcell wall components, aluminum silicate, glucan, or other known binders.In another embodiment, the commercial package, feed additive, additivefor the drinking water of the animal, or feed composition can furthercomprise inorganic/organic binders, essential oils, and/or organicacids.

In yet other embodiments, the commercial package, feed additive,additive for the drinking water of the animal, or feed compositioncomprising Bacillus strain 101, and/or Bacillus strain 235, and/orBacillus strain 77, and/or Bacillus strain 177, and/or Bacillus strain102 is in a container for commercial use. In various embodiments thecontainer can be, for example, a bag (e.g., a 20-pound bag, a 50-poundbag, a 2-ounce bag, a 1-pound bag, or a 1-kilogram bag), a pouch, adrum, a bottle, or a box. In illustrative aspects, the container for thecommercial package, feed additive, additive for the drinking water ofthe animal, or feed composition comprising Bacillus strain 101, and/orBacillus strain 235, and/or Bacillus strain 77, and/or Bacillus strain177, and/or Bacillus strain 102 can comprise plastic, metal, foil,paper, fiber, or cardboard (e.g., a plastic pail, a paper bag, a foilbag, a fiber drum, etc.). The commercial package, feed additive,additive for the drinking water of the animal, or feed composition canfurther comprise instructions for use of one or more of the Bacillusstrains.

The following examples are for illustrative purposes only. The examplesare non-limiting, and are not intended to limit the invention in anyway.

Example 1 Enzymatic Activity Screening

This example describes the use of plate media screening methods todetect enzymatic activity in DFM strains 101 and 235. Enzyme assay mediaplates were prepared by supplementing tryptic soy agar with between 0.5%and 1% of various substrates, including polysaccharides (corn starch,carboxymethylcellulose, or xylan), proteins (casein), and lipids(tributyrin). Bacillus strains of interest (including DFM strains 101and 235) obtained from fresh overnight cultures were spotted onto plates(5 μL) and incubated at various temperatures for up to 48 hours. Forprotein and lipid agar plates, zones of clearing around enzyme-producingcolonies were visible without further treatment.Polysaccharide-containing plates were stained with Gram's iodine for 1minute to visualize zones of clearing. ZOC diameters and colonydiameters were measured, but yes/no scores of activity were used forscreening purposes. DFM strains 101 and 235 were both observed to bepositive for protease, amylase, and xylanase activity after 48 hours andstrong positives on tributyrin agar (lipase) were observed.

The Bacillus strains described herein have enzymatic activity includingbut not limited to amylase, protease, xylanase and lipase.

Example 2 Antimicrobial Screening with the Cross-Streak Method

For facultative anaerobic pathogens tested (E. coli) the followingprotocol was used. Bacillus strains of interest (101 and 235) wereinoculated from frozen glycerol stocks in a single 1 cm wide linearstreak down the center of modified BHI agar plates. Bacillus-streakedplates were inoculated aerobically for 24 hours at 37° C., until a heavystreak of growth was present. The organism to be tested forsusceptibility, E. coli, was streaked in lines perpendicular towards theBacillus streak (up to 1 mm) in a biological safety cabinet andincubated at 37° C. for 24 hours under aerobic conditions. No more than5 staggered cross streaks per plate were used. The plates were incubatedand then examined for zones of inhibition around the initial Bacillusstreak, and the width of each zone of inhibition was measured inmillimeters. MDG strains 101 and 235 consistently inhibited E. coli. Todate, 58 different pathogenic E. coli strains were tested forsusceptibility to strains 101 and 43 different strains forsusceptibility to 235. These strains vary numerous characteristicsincluding toxin presence and quantity, colony morphology on SMAC agar,hemolysin production, and specimen location around the U.S. Strain 101successfully inhibited 46 of 58 E. coli isolates when it was grown bothaerobically and anaerobically while strain 235 successfully inhibited 42of 43 E. coli isolates when grown both aerobically and anaerobically.

Example 3 Culture Growth of DFM Strains 101 and 235

In the instant example, growth of DFM strains 101 and 235 can beachieved by culturing. On a small scale, TSB or nutrient broth can beutilized to culture DFM strains 101 and 235.

Agar medium may be produced using 23 grams Nutrient Agar (BD 213000) and1000 ml of DI water, followed by autoclaving at 121° C. Broth medium maybe produced using 8.0 grams of Nutrient Broth (BD 234000) and 1000 ml ofDI water, followed by autoclaving at 121° C.

For culturing DFM strain 101, a pure culture of DFM strain 101 wasstreaked on a nutrient agar plate and allowed to grow for 48 hours at32° C. Thereafter, a single colony was inoculated in nutrient brothmedium. The single colony was incubated at 37° C. and at 230-240 rpm,for 16 to 24 hours. Finally, the culture was streaked on a nutrient agarplate to check morphology.

For culturing DFM strain 235, a pure culture of DFM strain 235 wasstreaked on a nutrient agar plate and allowed to grow for 48 hours at32° C. Thereafter, a single colony was inoculated in nutrient brothmedium. The single colony was incubated at 37° C. and at 230-240 rpm,for 16 to 24 hours. Finally, the culture was streaked on a nutrient agarplate to check morphology.

Example 4 Antimicrobial Screening with the Cross-Streak Method

For E. coli tested, the protocol described in Example 2 was used. MGLstrains 77, 177, and 102 consistently inhibited E. coli. To date, 15different pathogenic E. coli strains were tested for susceptibility tostrains 77, 177, and 102. Strain 77 successfully inhibited 11 of 15 E.coli isolates when it was grown aerobically, strain 177 successfullyinhibited 12 of 15 E. coli isolates when it was grown aerobically, andstrain 102 successfully inhibited 15 of 15 E. coli isolates when it wasgrown aerobically (see Table 1 below).

TABLE 1 102 77 177 K 1.8 + + + (Beta) K 2.4 (beta) + + − K2.5(beta) + + + J 2.5 (beta) + + + K1.7 (beta) + + + K1.9 (beta) + − +K1.10 (beta) + − − K2.8 (beta) + + + K1.8 (beta) + + + J 1.2 (beta) + −− 1A (α) + − + 1B (α) + + + 1C (α) + + + 1D (α) + + + 1E (α) + + +

What is claimed is:
 1. A nutrient composition comprising an isolatedBacillus strain selected from the group consisting of Bacillus strain101 (NRRL No. B-67218), Bacillus strain 235 (NRRL No. B-67219), Bacillusstrain 77 (NRRL No. B-67274), Bacillus strain 177 (NRRL No. B-67275),Bacillus strain 102 (NRRL No. B-67276), a strain having all of theidentifying characteristics of Bacillus strain 101 (NRRL No. B-67218), astrain having all of the identifying characteristics of Bacillus strain235 (NRRL No. B-67219), a strain having all of the identifyingcharacteristics of Bacillus strain 77 (NRRL No. B-67274), a strainhaving all of the identifying characteristics of Bacillus strain 177(NRRL No. B-67275), a strain having all of the identifyingcharacteristics of Bacillus strain 102 (NRRL No. B-67276), andcombinations thereof, wherein the strain(s) are in a form selected fromthe group consisting of a pellet, a gel, a freeze-dried composition, atop-dressing, a liquid, and a dried powder.
 2. The composition of claim1 wherein the composition further comprises a carrier.
 3. Thecomposition of claim 1 wherein the composition further comprises anexogenously added nutrient component selected from the group consistingof a vitamin, an antibiotic, an enzyme, a water-soluble orwater-insoluble monosaccharide, disaccharide, or polysaccharide, a fat,phosphorous, sodium bicarbonate, limestone, calcium, sodium, sulfur,magnesium, potassium, copper, iron, manganese, zinc, fish oil, raw seed,an antioxidant, and a starch.
 4. The composition of claim 1 furthercomprising a binder wherein the binder is selected from the groupconsisting of clay, yeast cell wall components, aluminum silicate,glucan, and combinations thereof.
 5. The composition of claim 1 in theform of a commercial package.
 6. The composition of claim 5 wherein thecommercial package comprises a container selected from the groupconsisting of a bag, a pouch, a drum, a bottle, and a box.
 7. Thecomposition of claim 2 wherein the carrier is selected from the groupconsisting of a bran, rice hulls, a salt, mineral oil, a dextrin, whey,sugar, limestone, dried starch, sodium silico aluminate, vegetable oil,and combinations thereof.
 8. The composition of claim 3 wherein theexogenously added component is an enzyme.
 9. The composition of claim 8wherein the enzyme is selected from the group consisting of agalactosidase, a protease, a lipase, an amylase, a hemicellulase, anarabinoxylanase, a xylanase, a cellulase, an NSPase, a phytase, andcombinations thereof.